Bacteriophage control of Campylobacters in retail poultry
Food-borne disease continues to be a major cause of human morbidity and mortality. During the past few decades, Campylobacter jejuni has ascended to become the greatest cause of bacterial enteric disease worldwide. Anecdotal evidence suggests the majority of human campylobacteriosis in industrialised countries is caused by the consumption of undercooked chicken. Campylobacter continues to frustrate current control strategies throughout the food chain and in 2001 was responsible for over 56, 000 cases of food poisoning in the U.K. alone. The work presented in this thesis examined the potential of host-specific bacteriophage as a novel measure to control the population of Campylobacter in poultry production. Several surveys in this thesis revealed that campylobacters and their bacteriophage permeate the entire poultry meat supply chain, from chickens in the broiler house to packaged retail products. Characterisation of the bacteriophage recovered from such sources showed that retail poultry isolates exhibited greater similarities in host range than those originating from broiler houses, implying poultry processing selected for a subpopulation of phage. Additionally, broiler chickens harbouring bacteriophage in their gastrointestinal tract generally contained fewer campylobacters. All of the phage isolates studied belonged to the Myoviridae virus family as they possessed dsDNA genomes encapsulated in an icosahedral head with a rigid, contractile tail. Fragments of the phage genomes exhibited significant sequence homology with a number of genes involved in DNA replication from phage T4. Studies of the attachment and replication of the phage isolates in vitro suggested that adsorption to the host cell was efficient but the burst size was low (˂10 virions per cell). Campylobacter jejuni was found to produce membrane vesicles but these did not significantly affect bacteriophage replication in vitro. A series of trials using 'phage therapy' in broiler chickens revealed that Campylobacter colonisation can be reduced by ≥log[subscript]10 8.0 cfu g[superscript]-1 caecal contents by dosing with specific bacteriophage. However, both the timing and extent of the reduction in Campylobacter colonisation showed considerable variation. Additionally, the ability of bacteriophage to infect their host in vitro was not a reliable indicator of their efficacy in vivo. The direct application of bacteriophage to the surface of chicken skin artificially contaminated with Campylobacter led to a significant reduction in the number of recoverable host cells. Host resistance to bacteriophage infection was not detected in either the in vivo trials or when recovering Campylobacter cells from chicken skin treated with phage. The work presented in this thesis demonstrates that bacteriophage have considerable potential in the control of Campylobacter in poultry production. They already appear to constitute a limiting factor in Campylobacter colonisation of the chicken gastrointestinal tract and can be detected with their host on retail products. However, further research is required to fully realise their potential and optimising the timing, level and type of bacteriophage used in dosing will be important for their efficacy in vivo.